CN115990172A - Application of pentagalloylglucose in preparation of antidepressant drugs - Google Patents

Abstract

The invention provides an application of pentagalloylglucose in preparing antidepressant drugs, belonging to the technical field of biological medicines. The invention discovers that pentagalloylglucose can improve the depression-like behavior of the mouse with the deficiency of the pleasure of the CUMS, can improve the behavior destimacy state of the mouse, can increase the exploration behavior of the CUMS mouse, namely the pentagalloylglucose has the effect of improving the depression-like behavior of the mouse; meanwhile, the level of LC3B, beclin1 and p62 of the colon of the mouse can be obviously improved by 15mg/kg of pentagalloylglucose, and the level of LC3B, beclin1 and p62 in the forehead cortex of the mouse can be obviously improved by 30mg/kg of pentagalloylglucose, namely, autophagy of the colon and the forehead cortex of the mouse can be enhanced by the pentagalloylglucose. The invention discovers the antidepressant effect and the action mechanism of the pentagalloylglucose for the first time, and has great significance for treating depression.

Description

Application of pentagalloylglucose in preparation of antidepressant drugs

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to application of pentagalloylglucose in preparation of antidepressant drugs.

Background

Depression is now the most common psychological disorder, which is defined as a serious mood disorder according to the classification standard of the american handbook of diagnosis and statistics of mental disorders, and patients often show core symptoms of sadness, despair, lack of pleasure, discomfort and the like with accompanying cognitive and somatic discomfort, with a significant decrease in the aggressiveness to daily life. Depression has become one of the most common mental disorders and leading causes of disability. The prevalence of depression has grown year by year, and about 3.5 million depressed patients are reported worldwide.

At present, most of the drug treatments are mainly western medicines which are chemically synthesized, such as serotonin reuptake inhibitors, serotonin and norepinephrine dual reuptake inhibitors, and the like. The main effects of these agents are closely related to increasing synaptic cleft neurotransmitter levels. However, these chemical synthesis antidepressants generally have the disadvantages of slow effect, long period and large side effect, while the traditional Chinese medicine has the advantages of small side effect, abundant sources, wide targets and the like, so that the application of the traditional Chinese medicine in the aspect of treating depression is increasingly wide, and therefore, more traditional Chinese medicines capable of treating depression are necessary to be developed.

1,2,3,4, 6-O-pentagalloylglucose is a natural polyphenol compound which exists in plants such as paeonia lactiflora, peony, pelargonium and the like, has various biological activities including antioxidant, anti-inflammatory, cytoprotective, antiviral and other biological activities, and has been proved to play a certain role in a plurality of malignant tumor diseases such as breast cancer, prostate cancer, liver cancer and the like. PGG has been reported to induce autophagy in diseases such as prostate cancer and nasopharyngeal cancer, and to enhance the intrinsic antiviral activity of the body by enhancing autophagosome formation. However, in the nervous system and depression diseases, no report is found.

Disclosure of Invention

In order to solve the technical problems, the invention provides application of pentagalloylglucose in preparing antidepressant drugs. The invention discovers that pentagalloylglucose can improve the depression-like behavior of the mouse with the deficiency of the pleasure of the CUMS, can improve the behavior destimacy state of the mouse, can increase the exploration behavior of the CUMS mouse, namely the pentagalloylglucose has the effect of improving the depression-like behavior of the mouse; meanwhile, the level of LC3B, beclin1 and p62 of the colon of the mouse can be obviously improved by 15mg/kg of pentagalloylglucose, and the level of LC3B, beclin1 and p62 in the forehead cortex of the mouse can be obviously improved by 30mg/kg of pentagalloylglucose, namely, autophagy of the colon and the forehead cortex of the mouse can be enhanced by the pentagalloylglucose. The invention discovers the antidepressant effect and the action mechanism of the pentagalloylglucose for the first time, and has great significance for treating depression.

In order to achieve the above purpose, the present invention adopts the following technical scheme:

the invention provides an application of pentagalloylglucose in preparing antidepressant drugs, wherein the drug effect dosage of the antidepressant drugs is 10-40 mg of pentagalloylglucose per kilogram of body weight per day.

Preferably, the effective dosage of the antidepressant is pentagalloylglucose of 15-30 mg per kilogram of body weight per day.

Preferably, the pentagalloylglucose is the only active ingredient of the antidepressant.

Preferably, the antidepressant comprises at least one of a pharmaceutically acceptable salt, a co-crystal, a stereoisomer, a prodrug, a solvate, and a metabolite of pentagalloylglucose.

Preferably, the antidepressant further comprises pharmaceutically acceptable excipients.

Preferably, the auxiliary materials comprise at least one of diluents, excipients, fillers, binders, wetting agents, absorption promoters, surfactants, lubricants, stabilizers, flavoring agents, sweeteners, pigments which are conventional in the pharmaceutical field.

Preferably, the antidepressant can be prepared into any pharmaceutically acceptable dosage form, wherein the dosage form comprises any one of emulsion, cream, pill, dripping pill, capsule, granule, powder, paint, cataplasm, spray, oral liquid, decoction, injection, sustained release preparation or controlled release preparation.

Compared with the prior art, the invention has the following technical effects:

(1) The invention discovers that pentagalloylglucose can improve the depression-like behavior of CUMS mice caused by the lack of pleasure, can also improve the behavior destimation state of the mice, and can also increase the exploration behavior of the CUMS mice, which indicates that pentagalloylglucose can improve the depression-like behavior of the mice.

(2) Treatment with CUMS altered the expression of autophagy-related signaling molecules, including the levels of LC3B, beclin1 and p62, whereas pentagalloylglucose treatment reversed these changes. Specifically, the 15mg/kg pentagalloylglucose treated group significantly elevated the levels of LC3B, beclin1 and p62 in the mouse colon, whereas the 30mg/kg pentagalloylglucose treated group failed to elevate the levels of Beclin1 and p62 in the mouse colon, but the 30mg/kg pentagalloylglucose treated group significantly increased the levels of LC3B, beclin1 and p62 in the mouse forehead cortex. Indicating that pentagalloylglucose enhances autophagy in the colon and prefrontal cortex of mice.

(3) The invention discovers the antidepressant effect and the corresponding action mechanism of the pentagalloylglucose for the first time, and has very important significance for treating depression.

Drawings

FIG. 1 shows the results of the behavioural test in example 4 of the present invention, wherein A is the sugar water preference test result, B is the tail suspension test result, and C is the open field test result;

FIG. 2 shows the results of autophagy marker level detection in example 5 of the present invention, wherein A-C is the level of mouse colon LC3B, beclin, p62, and D-F is the level of mouse forehead cortex LC3B, beclin, p 62.

Detailed Description

The following examples are illustrative of the invention and are not intended to limit the scope of the invention. Modifications and substitutions to methods, procedures, or conditions of the present invention without departing from the spirit and nature of the invention are intended to be within the scope of the present invention.

The technical scheme of the invention is further elaborated in the following in conjunction with examples.

Example 1 grouping and treatment of animals

38 healthy male C57BL/6N SPF mice are selected, are bred in a medical animal laboratory with the weight of 20+/-5 g, and are subjected to an adaptive feeding process for 7 days before the beginning of a formal experiment. Feeding conditions: room temperature 21±2 ℃, relative moderate 30-40%, light brightness 12 hours (7:00-19:00, dark 19:00-7:00). Mice were randomized into 5 groups, control, chronic unpredictable Wen Heying stress depression model (CUMS), fluoxetine (FLX), pentagalloylglucose low dose (PGG 15 mg/kg), pentagalloylglucose high dose (PGG 30 mg/kg). Animals in the control group were kept normally without stimulation; model, FLX, and PGG animals were subjected to chronic unpredictable stress patterns for 8 weeks, while FLX and PGG groups received 3 weeks of treatment.

EXAMPLE 2 preparation of the medicament

PGG (99.61% purity, CAS number 14937-32-7) was purchased from Chengdu Maddson technology Co., ltd (China) (lot number RP 210608). PGG received a dose of 15mg/kg, 30mg/kg of 1,2,3,4, 6-0-pentagalloylglucose, respectively, once daily by the intragastric method. The PGG solution was prepared as follows: 84mg and 168mg of PGG were dissolved in pure water, respectively, and the final concentrations of PGG were 1.5mg/mL and 3mg/mL.

Fluoxetine (FLX) was purchased from beijing wokawa biotechnology limited (beijing, china) (lot OKA 20210408). FLX group received 20mg/kg of fluoxetine, administered once daily by gavage. The FLX solution was prepared as follows: 112mg of FLX was dissolved in pure water, and the final concentration of FLX was 2mg/mL.

EXAMPLE 3 preparation of CUMS model

The method of inducing CUMS is to alternately expose to a pressure source every day for 8 consecutive weeks. The stimulation mode comprises restraint for 3 hours, illumination over night, fasted for 24 hours, water forbidden for 24 hours, ice water swimming at 4-8 ℃ and tail clamping for 2 minutes, the stimulation is randomly arranged in one week, one kind of stimulation is daily, and the same stimulation can not continuously occur for 8 weeks. And carrying out a syrup preference experiment, a tail suspension experiment and an open field experiment before death of the animal broken end.

Example 4 behavioural test

1 syrup preference experiment (SPT)

The day before the start of the experiment, animals were trained to adapt to sugar-containing drinking water and two water bottles containing 1% sucrose were given to each mouse for 24 hours. Within 24 hours of the next day, a bottle of pure water and 1% sucrose was placed in each cage, and after 24 hours of fasting and water-deprivation on the third day, each mouse was given one bottle of pure water and one bottle of 1% sucrose at the same time. After one hour, the volumes of pure water and sucrose solution remaining were recorded. The animal's preference (%) for sugar water was calculated as follows.

Animal preference rate for sugar water (%) =sugar water intake (g)/total intake (g).

2 tail suspension experiment (TST)

Each mouse was suspended on a horizontal bar 50 cm from the ground, and the tail was fixed with tape. The entire experimental procedure lasted 6 minutes. The activity of each mouse was recorded over the last 4 minutes. Immobility time (S) is defined as the time required for a mouse to give up struggling and remain completely immobile.

3 open field experiment (OFT)

Each mouse was allowed to stand in the behavioral control chamber for 10 minutes for adaptation, then moved to the central zone, camera shooting was started and timing was performed, and the behavior of the mouse was observed for 5 minutes. Immediately after each experiment, the box was rinsed with 75% alcohol. The OFT was performed using internationally recognized behavioral analysis software (EthoVision software analysis System, noldus information technology Co., leesburg, va., USA).

The experimental data of the three experiments are statistically analyzed by SPSS25 software, the comparison among groups is single-factor analysis of variance (ANOVA), the experimental data is expressed by mean plus-minus standard deviation (x+ -s), and when P is less than 0.05, the difference is considered to have statistical significance. The sugar water preference experiment result is shown in figure 1A, the tail suspension experiment result is shown in figure 1B, and the open field experiment result is shown in figure 1C.

As can be seen from fig. 1A: compared with the control group, the sugar water preference rate of the CUMS group is significantly reduced (P < 0.001); the increased syrup bias rate (P < 0.01) in FLX and PGG groups compared to CUMS group suggests that PGG can improve depression-like behavior of CUMS mice with loss of pleasure.

As can be seen from fig. 1B: the CUMS group had a significant rise in immobility time (P < 0.001) compared to the control group; compared with the CUMS group, the FLX group and the PGG group have reduced immobility time (P < 0.05), which suggests that PGG can improve the behavior destimation state of mice.

As can be seen from fig. 1C: compared with the control group, the CUMS group has reduced residence time in the central area of open field (P < 0.05); the increase in residence time (P < 0.05) in the open field central region for the FLX and PGG groups compared to the CUMS group suggests that PGG may increase exploratory behavior in CUMS mice.

The above behavioral tests demonstrate that PGG uptake can alleviate CUMS-induced depressive-like and anxiety-like behaviors in mice.

Example 5 autophagy marker level detection

The PGG treatment group was tested by RT-qPCR at a low dose of 15mg/kg and a high dose of 30mg/kg to detect the levels of three commonly used autophagy markers LC3B, beclin1 and p62 in the colon and prefrontal cortex of mice after 3 weeks of treatment with PGG, respectively, to investigate autophagy under CUMS-induced depression-like conditions.

Total RNA was extracted from colon and forehead cortex homogenate tissues of mice using Soxhaust Total RNA extraction kit (Beijing) according to the manufacturer's instructions. Quantification was performed on a Bio-rad Cx96 detection system (American biological Radar) using reverse transcription kit and SYBR premix qPCR kit (Ai Kerui) and gene specific primers, wherein the primers used for PCR are shown in Table 1.

TABLE 1LC3B, beclin, p62 primer sequences

Each cDNA was tested in triplicate. The conditions of the heat source were 95℃for 30 minutes, 95℃for 5 seconds, 60℃for 30 seconds, and 40 cycles of amplification. Relative quantitative normalization of PCR products, GAPDH served as an internal reference.

The experimental data are also statistically analyzed by SPSS25 software, the group comparison is by single-factor analysis of variance (ANOVA), the experimental data are expressed by mean plus or minus standard deviation (x+ -s), and when P <0.05, the difference is considered to have statistical significance. The results of the test are shown in FIG. 2, wherein FIG. 2A-FIG. 2C show the levels of mouse colon LC3B, beclin1, p62, and FIG. 2D-FIG. 2F show the levels of mouse forehead cortex LC3B, beclin1, p 62.

The results show that: treatment with CUMS altered the expression of autophagy-related signal molecules, including the levels of LC3B, beclin1 and p62, while treatment with PGG reversed these changes. As shown in fig. 2A-2C, the 15mg/kg pgg treated group significantly increased the levels of mouse colon LC3B, beclin1 and p62, but the 30mg/kg pgg treated group failed to increase the levels of mouse colon Beclin1 and p62, compared to the control group. However, as can be seen from FIGS. 2D-2F, the levels of LC3B, beclin1 and p62 were significantly increased in the prefrontal cortex of mice in the 30mg/kg PGG treated group. These data indicate that PGG enhances autophagy in the colon and prefrontal cortex of mice.

The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.

Claims (7)

1. The application of pentagalloylglucose in preparing antidepressant is characterized in that the pharmacodynamic dose of the antidepressant is 10-40 mg of pentagalloylglucose per kilogram of body weight per day.

2. The use according to claim 1, wherein the antidepressant is administered in a dose of from 15 to 30mg pentagalloylglucose per kg body weight per day.

3. The use according to any one of claims 1 to 2, characterized in that pentagalloylglucose is used as the sole active ingredient of the antidepressant drug.

4. The use according to any one of claims 1-2, wherein the antidepressant comprises at least one of a pharmaceutically acceptable salt, co-crystal, stereoisomer, prodrug, solvate, metabolite of pentagalloylglucose.

5. The use according to any one of claims 1 to 2, wherein the antidepressant further comprises a pharmaceutically acceptable adjuvant.

6. The use according to claim 5, wherein the auxiliary materials comprise at least one of diluents, excipients, fillers, binders, humectants, absorption enhancers, surfactants, lubricants, stabilizers, flavoring agents, sweeteners, pigments, which are conventional in the pharmaceutical field.

7. The use according to any one of claims 1 to 2, wherein the antidepressant is formulated in any pharmaceutically acceptable form, including any one of an emulsion, a cream, a pill, a drop pill, a capsule, a granule, a powder, a paint, a cataplasm, a spray, an oral liquid, a decoction, an injection, a sustained release preparation or a controlled release preparation.

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